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Open AccessArticle

A New Cytotoxicity Assay for Brevetoxins Using Fluorescence Microscopy

UNCW Center for Marine Science, 5600 Marvin K Moss Lane, Wilmington, NC 28409, USA
Author to whom correspondence should be addressed.
Mar. Drugs 2014, 12(9), 4868-4882;
Received: 1 August 2014 / Revised: 1 September 2014 / Accepted: 4 September 2014 / Published: 23 September 2014
(This article belongs to the Special Issue Marine Dinoflagellates)
Brevetoxins are a family of ladder-framed polyether toxins produced during blooms of the marine dinoflagellate, Karenia brevis. Consumption of shellfish or finfish exposed to brevetoxins can lead to the development of neurotoxic shellfish poisoning. The toxic effects of brevetoxins are believed to be due to the activation of voltage-sensitive sodium channels in cell membranes. The traditional cytotoxicity assay for detection of brevetoxins uses the Neuro-2A cell line, which must first be treated with the neurotoxins, ouabain and veratridine, in order to become sensitive to brevetoxins. In this study, we demonstrate several drawbacks of the Neuro-2A assay, which include variability for the EC50 values for brevetoxin and non-linear triphasic dose response curves. Ouabain/ veratridine-treated Neuro-2A cells do not show a typical sigmoidal dose response curve in response to brevetoxin, but rather, have a polynomial shaped curve, which makes calculating EC50 values highly variable. We describe a new fluorescence live cell imaging model, which allows for accurate calculation of cytotoxicity via nuclear staining and additional measurement of other viability parameters depending on which aspect of the cell is stained. In addition, the SJCRH30 cell line shows promise as an alternative to Neuro-2A cells for testing brevetoxins without the need for ouabain and veratridine. View Full-Text
Keywords: brevetoxins; cytotoxicity assay; triphasic response; hormesis; Neuro-2A; SJCRH30 brevetoxins; cytotoxicity assay; triphasic response; hormesis; Neuro-2A; SJCRH30
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MDPI and ACS Style

McCall, J.R.; Elliott, E.A.; Bourdelais, A.J. A New Cytotoxicity Assay for Brevetoxins Using Fluorescence Microscopy. Mar. Drugs 2014, 12, 4868-4882.

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